Intracellular acidification induced by membrane depolarization in rat hippocampal slices: roles of intracellular Ca2+ and glycolysis.
To elucidate the mechanism of pHi changes induced by membrane depolarization, the variations in pHi and [Ca2+]i induced by a number of depolarizing agents, including high K+, veratridine, N-methyl-D-aspartate (NMDA) and ouabain, were investigated in rat hippocampal slices by the fluorophotometrical technique using BCECF or fura-2. All of these depolarizing agents elicited a decrease in pHi and an elevation of intracellular calcium ([Ca2+]i) in the CA1 pyramidal cell layer. The increases in [Ca2+]i caused by the depolarizing agents almost completely disappeared in the absence of Ca2+ (0 mM Ca2+ with 1 mM EGTA). In Ca2+ free media, pHi acid shifts produced by high K+, veratridine or NMDA were attenuated by 10-25%, and those produced by ouabain decreased by 50%. Glucose-substitution with equimolar amounts of pyruvate suppressed by two-thirds the pHi acid shifts induced by both high K+ and NMDA. Furthermore, lactate contents were significantly increased in hippocampal slices by exposure to high K+, veratridine or NMDA but not by ouabain. These results suggest that the intracellular acidification produced by these depolarizing agents, with the exception of ouabain, is mainly due to lactate accumulation which may occur as a result of accelerated glycolysis mediated by increased Na+-K+ ATPase activity. A Ca2+-dependent process may also contribute to the intracellular acidification induced by membrane depolarization. Since an increase in H+ concentration can attenuate neuronal activity, glycolytic acid production induced by membrane depolarization may contribute to the mechanism that prevents excessive neuronal excitation.
Zhan
R Z
RZ
Department of Anesthesiology, Niigata University School of Medicine, Asahimachi-dori, Japan.
Fujiwara
N
N
Tanaka
E
E
Shimoji
K
K
D000143:Acids; D000818:Animals; D002118:Calcium / Q000502:physiology*; D005452:Fluoresceins; D005456:Fluorescent Dyes; D015590:Fluorophotometry; D016257:Fura-2; D006019:Glycolysis / Q000502:physiology*; D006624:Hippocampus / Q000502:physiology*; D006863:Hydrogen-Ion Concentration; D066298:In Vitro Techniques; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D010042:Ouabain / Q000494:pharmacology*; D011188:Potassium / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D014701:Veratridine / Q000494:pharmacology*
D000143:Acids; D005452:Fluoresceins; D005456:Fluorescent Dyes; D010042:Ouabain; D014701:Veratridine; C043829:2',7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; D011188:Potassium; D002118:Calcium; D016257:Fura-2